寻常型银屑病患者外周血CD4~+CD25~+及CD8~+CD25~+细胞的检测

目的研究进展期寻常型银屑病患者外周血CD4+CD25+和CD8+CD25+调节性T细胞的数量变化及其在银屑病免疫病理学发病机制中的作用。方法应用流式细胞术对进展期寻常型银屑病患者外周血CD4+CD25+和CD8+CD25+调节性T细胞进行检测。结果进展期寻常型银屑病外周血CD4+CD25+细胞及CD8+CD25+调节性T细胞数量与正常对照组相比,均显著降低(P<0.05,P<0.005),而CD4+CD25+/CD8+CD25+比值无显著性差异(P>0.05)。结论寻常型银屑病的发病与CD4+CD25+和CD8+CD25+调节性T细胞的同步降低有关,与二者的比值无关。     

银屑病患者造血细胞定向分化的CD4+ CD25+调节性T细胞活性研究

目的:探讨造血细胞与银屑病患者CD4 CD25 节性T细胞活性异常的关系.方法:在胸腺基质细胞支持下,将骨髓CD34 细胞向T细胞定向分化,免疫磁珠法分离CD4 CD25 调节性T细胞,分别在自然增殖状态及链球菌超抗原刺激下,采用四甲基偶氮唑蓝(MTT)法和ELISA法测定定向分化的CD4 CD25 T细胞增殖活性及分泌细胞因子水平.结果:在自然增殖状态下,银屑病患者CD34 细胞定向分化的CD4 CD25 T细胞显示与正常对照相似的增殖活性及白介素(IL)-2、IL-4、IL-8、IL-10、干扰素(IFN)-γ等细胞因子分泌水平,然而,在受到A族链球菌超抗原刺激后,银屑病患者定向分化的CD4 CD25 T细胞较正常对照显示相对低的增殖活性及分泌IL-2、IL-10能力.结论:银屑病患者CD34 细胞定向分化的CD4 CD25 T细胞功能异常,提示银屑病患者调节性T细胞功能异常可能源自遗传背景决定的骨髓造血细胞活性.     

Functional characterization of CD4+CD25+ regulatory T cells differentiated in vitro from bone marrow-derived haematopoietic cells of psoriasis patients with a family history of the disorder

BACKGROUND: In psoriasis CD4+CD25+ regulatory T cells are functionally deficient. The imbalance between regulatory and effector T-cell functions is important for inducing psoriasis. It is reasonable to speculate that the dysfunctional activity of CD4+CD25+ regulatory cells may originate partly from the abnormal haematopoietic cells determined mainly by genetic background. OBJECTIVES: To test the hypothesis that haematopoietic stem cells are responsible for dysfunctional CD4+CD25+ regulatory cells in psoriasis. METHODS: Bone marrow-derived CD34+ haematopoietic cells from patients with psoriasis (with a family history of psoriasis) and from normal controls were differentiated into T cells in vitro. CD4+CD25+ T cells were isolated by an immunomagnetic bead method, and proliferation activity and capacity for cytokine secretion were determined. Furthermore, the ability of CD4+CD25+ T cells to suppress the proliferative responses of allogeneous peripheral blood CD4+CD25- effector T cells was assessed in vitro. RESULTS: The differentiated CD4+CD25+ T cells of psoriatic origin showed similar characteristics to those of normal volunteers, including proliferation activity and secretion profile of the cytokines interleukin (IL)-2, IL-4, IL-8, IL-10 and interferon (IFN)-gamma. However, proliferation and secretion levels of the cytokines IL-2 and IL-10 for CD4+CD25+ cells of psoriatic CD34+ cell origin were significantly lower than those of normal controls in response to streptococcal superantigen (Strep-A). In particular, CD4+CD25+ T cells differentiated from psoriatic CD34+ cells were functionally insufficient to restrain effector T-cell proliferation. CONCLUSIONS: CD4+CD25+ T cells differentiated in vitro from haematopoietic cells of patients with psoriasis are impaired in regulatory function. The dysfunction of psoriatic CD4+CD25+ T cells may be due to inherent genetic programming passed down from bone marrow-derived haematopoietic cells.     

Persistent CMV infection correlates with disease activity and dominates the phenotype of peripheral CD8+ T cells in psoriasis

Background: Previously, we have reported a frequent association of active plaque psoriasis with inflammation‐mediated cytomegalovirus (CMV) reactivation. Objectives: This study aimed at characterizing the impact of CMV infection on psoriasis disease activity and peripheral cellular adaptive immune response. Patients/Methods: Twenty nine patients with active plaque psoriasis and 29 healthy controls were analysed for CMV‐serostatus, CMV‐antigenaemia, frequencies of peripheral CMV‐specific T cells and the immunophenotype of peripheral CD8+ T cells. Results: (i) Psoriasis severity was higher in CMV‐seropositive patients and positively correlated to the severity of CMV‐antigenaemia. (ii) In comparison to CMV‐seropositive healthy controls, CMV‐seropositive psoriasis patients showed a reduced frequency of circulating CMV‐specific T cells that increased under effective antipsoriatic therapy. (iii) The immunophenotype of peripheral CD8+ T cells was dominated by CMV‐seroprevalence. (iv) Selective analysis of CMV‐seronegative psoriasis patients revealed a strong expansion of a – probably early activated – CD8+ T‐cell population with the yet undescribed differentiation phenotype ‘CD45RA‐dim/CD11a‐dim’. Under effective antipsoriatic therapy this population decreased in parallel to an increase of effector differentiated CD8+ T cells. Conclusions: Taken together with our previous results of inflammation‐mediated CMV reactivation in psoriasis, our data support the concept of an interactive relationship between psoriasis and CMV infection which may be mediated by peripheral CD8+ T cells.     

银屑病患者骨髓CD34+细胞定向分化的T细胞对角质形成细胞增殖的影响

目的:探讨家族史阳性的银屑病患者骨髓CD34 细胞体外定向分化的T细胞对表皮角质形成细胞(KC)增殖调控蛋白表达的影响.方法:将银屑病患者骨髓CD34 细胞体外定向分化的T细胞经链球菌超抗原活化后与KC共培养,分别采用免疫组化法及ELISA法检测KC C-myc、bcl-xL、p53及Ki67蛋白表达及培养上清白介素(IL)-8、干扰素(IFN)-γ水平.结果:①受银屑病患者CD34 细胞定向分化T细胞作用的KC C-mye及Ki67蛋白表达与自然增殖组及正常人CD34 细胞定向分化的T细胞作用组相比显著增强(P＜0.05),而bcl-xL及p53蛋白表达3组间的差异无统计学意义(P＞0.05);②受正常人CD34 细胞定向分化T细胞作用的Kc C-myc、bcl-xL、p53及Ki67蛋白表达与自然增殖组比较差异亦无统计学意义(P＞0.05);③银屑病CD34 细胞定向分化的T细胞作用组培养上清IL-8及IFN-γ水平显著高于正常对照组(P＜0.01).结论:银屑病患者骨髓造血细胞定向分化的T细胞可影响KC增殖状态,显示类似银屑病外周血T细胞的活性特点.     

CD103+T细胞在银屑病皮损中的表达

目的：明确银屑病患者皮肤CD103+T细胞的表达及其与银屑病严重程度的关系。方法：免疫组化检测29例银屑病患者皮损和非皮损皮肤及6名健康对照皮肤中表皮及真皮CD103+T细胞的表达。计算银屑病患者PASI值。结果：CD103+T细胞主要在真皮表达。银屑病患者皮损和非皮损真皮中每个高倍视野CD103+T细胞百分率分别为（26.06%±11.72）%和（12.82±4.5）%（P<0.05）;健康人对照皮肤真皮内CD103+T细胞百分率为（7.47±1.3）%,明显低于银屑病非皮损区（P<0.05）。银屑病患者皮损中,CD103+T的表达与PASI值正相关（P<0.05）。 结论：真皮中CD103+T细胞可能与银屑病的发病及严重程度有关。     

Epidermal CD8+ T cells in chronic plaque psoriasis are Tc1 cells producing heterogeneous levels of interferon-gamma

The majority of epidermal CD8+ T cells in chronic plaque psoriasis are activated Tc1 cells producing interferon-gamma and no interleukin-4, a small proportion of which express NK-T receptors. To quantitate their level of cytokine production and characterize them further, CD8+ T cells were isolated from epidermal cell suspensions of lesional biopsies from 24 patients with chronic plaque psoriasis. T-cell lines (TCL) were established by culture of CD8+ T cells with feeders and IL-2 for 11 days and expansion with PHA. Ten TCL were stained for surface markers; 6 were cloned with PHA by limiting dilution. Interferon-gamma, interleukin-4 and interleukin-10 production was measured by ELISA after PMA/anti-CD3 activation of 15 TCL and 39 CD8+ T-cell clones. The 10 TCL stained were CD8alphabeta+ (93.3%), T-cell receptor-alphabeta+ (99.5%), costimulatory molecule CD28+ (90.1%), with a small CD8alphaalpha+ population (2.3%). No NK-T-cell receptor CD158a or CD158b expression was detected, whilst CD94 was expressed on 6.2% of cells in 6/9 TCL. All the TCL and 37/39 CD8+ T-cell clones produced interferon-gamma but no or minimal interleukin-4 or interleukin-10. The TCL produced a wide range of interferon-gamma levels (138 to 15,020 pg/ml). Clones from 3 patients showed low levels (60 to 1,410 pg/ml), from 2 patients high levels (6,105 to 43,040 pg/ml) and from 1 patient a wide range (405 to 36,010 pg/ml) of interferon-gamma production. Thus epidermal CD8+ Tc1 cells in chronic plaque psoriasis produce highly heterogeneous levels of interferon-gamma, which may reflect clinical diversity.     

银屑病患者体外系统血液固有免疫反应中CD4~+ CD25~+ T细胞及CD4~+ CD25~(high) T细胞的变化与意义

目的用体外系统血液固有免疫反应体系研究银屑病患者外周血CD4+ CD25+T细胞及CD4+ CD25highT细胞的变化,并探讨其在银屑病发病机制中的作用。方法将灭活大肠杆菌悬液(3×108/mL)0.2mL作为免疫原激活剂加入枸橼酸抗凝的全血细胞悬液0.2mL和血浆0.3mL中,37℃水浴1h,用流式细胞仪测定CD4+ CD25+T细胞及CD4+ CD25highT细胞的比例。结果在加入大肠杆菌的银屑病全血细胞组(银屑病实验组)CD4+ CD25highT细胞比例(1.88%)明显高于银屑病对照组(1.41%)(P﹤0.01),CD4+ CD25+T细胞及CD4+ CD25highT细胞的比例(16.86%,1.88%)明显低于加入大肠杆菌的健康实验组(24.26%,2.81%)(P﹤0.01);银屑病对照组CD4+ CD25+T细胞及CD4+ CD25highT细胞的比例(15.97%,1.41%)较健康对照组明显降低(21.75%,2.17%)(P﹤0.01);健康实验组CD4+ CD25+T细胞及CD4+ CD25highT细胞的比例群(24.26%,2.81%)均明显高于健康对照组(21.75%,2.17%),(P﹤0.05)。结论银屑病患者CD4+ CD25+T细胞及CD4+ CD25highT细胞比例降低,外来抗原刺激后比例升高,但与健康正常人存在差异。这可能与其复杂的系统免疫学发病机制相关。     

CD4~+CD25~+调节性T细胞与Th17细胞在寻常型银屑病中的表达

目的:检测外周血中CD4~+CD25~+调节性T细胞(简称Treg细胞)与Th17细胞在寻常型银屑病(PV)中的表达。方法:34例寻常型银屑病患者,18例正常人作为对照。采用流式细胞术检测外周血中Treg细胞、FOXP3~+Treg细胞及Th17细胞的表达水平。结果:PV患者外周血中Treg细胞比例和FOXP3~+Treg细胞比例分别为(3.68±1.22)%和(0.53±0.19)%,低于正常对照组的(6.63±1.00)%和(0.76±0.14)%(P0.05)。Th17细胞比例为(2.20±0.78)%,高于正常对照组的(0.65±0.22)%(P0.05)。PV患者外周血中Treg细胞的表达与FOXP3~+Treg细胞的表达呈正相关(r=0.563,P0.05),而Treg细胞的表达与Th17细胞的表达呈负相关(r=-0.522,P0.05);Treg细胞和Th17细胞二者与PASI评分无相关性,与病程亦无相关性。结论:Treg细胞表达的减少及Th17细胞应答的增强,可能是导致PV免疫失衡的重要原因。     

银屑病外周血CD4+CD25+Foxp3+调节性T细胞的表达

目的: 检测CD4+CD25+Foxp3+调节性T细胞在不同类型银屑病患者中的表达.方法: 应用流式细胞仪检测外周血中CD4+CD25+Foxp3+调节性T细胞的表达.结果: 红皮病型银屑病患者外周血中CD4+CD25+Foxp3+调节性T细胞明显高于其他类型银屑病和正常对照组(P<0.05);斑块状银屑病CD4+CD25+Foxp3+T细胞比例高于点滴状患者(P<0.05);脓疱型银屑病患者中脓疱存在患者调节性T细胞比例明显低于脓疱消退患者(P<0.05).结论: CD4+CD25+Foxp3+调节性T细胞通过抑制效应T细胞在银屑病的病情活动中可能发挥重要作用.     

CD+4 CD+25调节性T细胞在自身免疫病中的意义

阐明CD4 CD2 5调节性T细胞作用机制及其自身免疫病的关系。     

Epidermal CD8+ T cells reactive with group A streptococcal antigens in chronic plaque psoriasis

Chronic plaque psoriasis is a T cell mediated disease associated with group A streptococci (GAS). We have previously shown the presence of a psoriasis-specific dermal Th1 subset that recognizes GAS antigens. To assess whether GAS-reactive T cells are also present in lesional epidermis, fresh cell suspensions or T cell lines isolated from lesional epidermis of 33 psoriasis patients were stained for intracellular interferon-gamma after stimulation with GAS antigens. The patients were typed by PCR for HLA-DR7 and HLA-Cw6 expression. A subset of GAS-reactive CD8+ T cells (2.4% +/- 2.4) was found in 14/21 (67%) fresh cell suspensions. A smaller subset of GAS-reactive CD4+ T cells (0.9% +/- 0.9) was found in 13/21 (62%) fresh cell suspensions, which was expanded in the T cell lines. There was a significant inverse correlation between the proportions of GAS-reactive CD4+ and CD8+ T cells in the fresh suspensions (r = -0.48, P = 0.0277). The presence of GAS-reactive CD4+ or CD8+ T cells did not correlate with HLA-DR7 or HLA-Cw6 expression, respectively. This study has demonstrated GAS-reactive CD8+, and to a lesser extent CD4+, T cell subsets in psoriatic epidermis and provides further evidence that GAS antigens may play a role in the pathogenesis of chronic plaque psoriasis.     

Expression of dipeptidyl-peptidase IV (CD26) on CD8+ T cells is significantly decreased in patients with psoriasis vulgaris and atopic dermatitis

T cells play a major role in inflammatory skin disorders such as psoriasis vulgaris and atopic dermatitis. They are both active on the level of cell-to-cell interaction and by the secretion of pro-inflammatory mediators. CD26 is a lymphocyte membrane-associated dipeptidyl peptidase IV (DPP IV), which is able to inactivate chemokines such as RANTES or eotaxin by cleaving dipeptides from the NH2-terminus of proteins. We investigated the expression of CD26 on CD4+ and CD8+ peripheral blood T cells in patients with psoriasis and atopic dermatitis. In addition PASI and SCORAD as a measure of disease severity were determined in each patient at the time of blood drawing. Thirty patients with psoriasis, 15 with atopic dermatitis and 17 age- and sex-matched healthy persons were investigated by two-colour flow cytometry using epitope-specific monoclonal antibodies. Our results revealed, that there is a significant decrease (P<0.05) of CD26 expression on CD8+ T cells in both psoriasis (7.7%+/-3.3, mean and SD, n=30) and atopic dermatitis patients (7.9%+/-3.7, mean and SD, n=15) compared to the control population (11.58%+/-5.0, mean and SD, n=17). However, there was no correlation to disease severity as determined by PASI and SCORAD, respectively. Since CD26 can be regarded as an anti-inflammatory principle the decreased expression in psoriasis and atopic dermatitis patients may lead to a dysbalance in favour of pro-inflammatory mediators in both clinical conditions.     

SLE患者外周血中CD4＋CD25＋CD127^low/-标记的调节性T细胞与临床表现和实验室指标相关性研究

目的：探讨CD4＋CD25＋CD127low/-标记的调节性T细胞（Treg）在系统性红斑狼疮发病机制电的作用。方法：用流式细胞仪检测45例系统性红斑狼疮（SLE）患者和45例年龄、性别相匹配的健康志愿者外周血CD4＋T细胞中Treg的百分比,同时分析SLE患者外周血中的CD4＋CD25＋CD127low/-标记的Treg与其临床表现、实验室指标的相关性。结果：SLE患者外周血CD4＋T细胞中Treg百分比与健康对照组比较差异无统计学意义。在SLE患者中,Treg的百分比与抗核小体抗体呈正相关（r=0．299,P=0．046）,有光敏感的SLE患者中Treg较无光敏感组增高（P=0．017）,余均无统计学差异。结论：SLE患者外周血中以CD4＋CD25＋CD127low/-标记的Treg中可能因含有部分无免疫抑制活性的效应性T细胞而特异性差,因此可能不适合用于临床免疫抑制治疗。     

HIV/AIDS患者CD4~+ T细胞中CD25和CD8~+ T细胞中CD28的表达

目的探讨外周血CD4+ T细胞中CD25表达率和CD8+ T细胞中CD28表达率在HIV/AIDS患者发病中的作用。方法应用流式细胞仪荧光染色技术检测35例HIV/AIDS患者CD4+ T细胞中CD25表达和CD8+ T细胞中CD28的表达,以41名健康体检人员做对照。结果HIV/AIDS患者和健康对照组之间CD4+ T细胞中CD25表达率(27.51±4.23)%,(44.41±9.17)%,CD4+25+ T细胞占淋巴细胞的比例(2.00±1.42)%,(16.62±4.60)%,CD4+25- T细胞占淋巴细胞的比例(5.16±3.37)%,(21.03±6.19)%,CD8+ T细胞中CD28中的表达率(25.12±6.33)%,(44.24±8.61)%,CD8+28- T细胞占淋巴细胞的比例(36.85±8.98)%,(13.33±4.58)%的差异均有显著性(P<0.01),CD8+ 28+ T细胞占淋巴细胞的比例(12.31±4.14)%,(10.51±3.71)%差异无显著性(P>0.05)。结论HIV/AIDS患者CD25在CD4+ T细胞与CD28在CD8+ T细胞的表达率降低可能与HIV感染后引起的免疫缺陷有关,CD8+ CD28- T细胞的增加有助于促进HIV/AIDS患者的炎性反应和免疫激活。     

银屑病患者角质形成细胞对T淋巴细胞CD25、CD69表达的影响

目的 探讨银屑病患者角质形成细胞（KC）对T淋巴细胞CD25、CD69表达的影响。方法 分离10例银屑病患者KC，密度梯度离心法分离单一核细胞（PBMC）；流式细胞仪检测混合培养后T细胞活化标志CD25、CD69的表达。结果 银屑病患者皮损KC作用的自体外周血T细胞CD25、CD69表达水平分别与非皮损KC作用组及自体T细胞自然增殖组相比显著增高，银屑病非皮损KC+自体T细胞共培养组与自体T细胞自然增殖组相比，差异无统计学意义。银屑病皮损、非皮损KC作用的正常人T细胞CD25、CD69表达均显著高于正常人外周血T细胞自然增殖组，银屑病皮损KC+正常人T细胞共培养组与非皮损组相比，差异无统计学意义。结论 银屑病患者局部存在慢性炎症反应，可能是由于皮损KC免疫表型发生改变从而作为自身抗原，启动自身免疫反应。     

外周血及蜕膜组织中CD4~+ CD25~+调节性T细胞比例和炎症因子原因不明复发性流产的关系研究

目的:探究外周血及蜕膜组织中CD4~+CD25~+调节性T细胞比例以及相关炎症因子对原因不明复发性流产的影响。方法:选取2014年5月至2015年5月就诊于我院门诊的原因不明复发性流产(URSA)患者90例为观察组,并选取同期在妇产科门诊手术室的行人工流产手术的正常患者90例为对照组;分析两组患者的一般资料、外周血及蜕膜组织中CD4~+CD25~+调节性T细胞比例、调节性T细胞特异调节因子Foxp3和EBi3的表达以及血清炎症因子(包括TNF-α、IL-2、IL-6、IL-10)水平,研究其相关性。结果:与正常妊娠组相比,反复流产组外周血及蜕膜组织中Treg细胞比例明显降低,Foxp3和EBi3因子mRNA的表达水平明显降低,且无论URSA患者或者是正常对照组,蜕膜中各因子的表达均高于外周血中的表达,外周血TNF-α、IL-2水平明显升高,而IL-6和IL-10水平要明显降低,差异有统计学意义(P0.05)。结论:蜕膜中各因子的表达均高于外周血中的表达提示妊娠过程的主要免疫反应部位是"母-胎界面",Treg细胞比例、Treg细胞因子和Th2型因子IL-6、IL-10水平可能参与妊娠的维持,调控"母-胎界面"局部免疫耐受。对于复发性流产的发病过程具有一定的影响作用。     

活动性皮肌炎CD4+CD25+CD127lo/-调节性T细胞的检测

皮肌炎(DM)是一种自身免疫性结缔组织病.CD4+CD25+调节性T细胞(Treg)是存在于人胸腺和外周血中的一类具有免疫抑制作用的T细胞亚群,该细胞数量减少或者功能异常均可导致自身免疫性疾病的发生.转录因子Foxp3是Treg细胞的特征性检测分子.研究发现,表面标志CD127(IL-7αR)的表达与Foxp3呈负相关,可以代替Foxp3作为Treg细胞的检测标志[1].为研究Treg细胞在活动性DM患者发病机制中的作用,我们分别对17例活动性DM患者和正常人对照外周血中的CD4+CD25+、CD4+CD25+CD127lo/-T细胞的百分比进行检测,同时与患者血清肌酸激酶(CK)含量进行相关性分析.并对其中7例住院患者给予糖皮质激素联合免疫抑制剂治疗,观察治疗后10 d,20 d CD4+CD25+和CD4+CD25+CD127lo/-T细胞百分比的变化.